摘要
目的:探讨栀子多糖对脂多糖(LPS)诱导的心肌细胞炎症反应、凋亡的影响及分子机制。方法:100 ng/ml LPS处理H9C2细胞作为LPS组,正常培养的细胞作为对照组,采用终浓度为2.5、5、10μmol/L的栀子多糖和100 ng/ml LPS共同培养的细胞作为栀子多糖低、中、高浓度组。将H9C2细胞分为LPS+Experiment-H+anti-miR-NC组、LPS+Experiment-H+anti-miR-141-3p组、LPS+Experiment-H+pcDNA3.1组、LPS+Experiment-H+pcDNA3.1-KLF6组。ELISA检测IL-1β、TNF-α水平;Western blot检测Bcl-2相关X蛋白(Bax)、Krupple样转录因子6(KLF6)蛋白表达;流式细胞术检测细胞凋亡;RT-qPCR检测miR-141-3p表达;荧光素酶报告实验检测miR-141-3p和KLF6的靶向关系。结果:LPS诱导的心肌细胞中IL-1β、TNF-α水平、Bax表达、细胞凋亡率显著升高,miR-141-3p表达显著降低,KLF6表达显著升高(P<0.05)。栀子多糖处理后LPS诱导的心肌细胞中IL-1β、TNF-α水平、Bax表达、细胞凋亡率显著降低,miR-141-3p表达显著升高,KLF6表达显著降低(P<0.05)。抑制miR-141-3p表达和过表达KLF6逆转了栀子多糖对LPS作用的H9C2细胞凋亡和炎症因子的抑制作用。miR-141-3p可靶向调控KLF6表达。结论:栀子多糖可抑制LPS诱导的心肌细胞凋亡和炎症因子释放,对LPS诱导的心肌细胞损伤具有保护作用,其机制可能与miR-141-3p和KLF6有关。
Objective:To investigate effects of gardenia polysaccharide on lipopolysaccharide(LPS)-induced cardiomyocyte inflammatory response and apoptosis and its molecular mechanism.Methods:H9C2 cells treated with 100 ng/ml LPS were set as LPS group,and normal cultured cells were set as control group,cells co-cultured with final concentrations of 2.5,5,10 μmol/L gardenia polysaccharide and 100 ng/ml LPS were set as gardenia polysaccharide low,medium and high concentrations groups. H9C2 cells were divided into LPS+Experiment-H+anti-miR-NC group,LPS+Experiment-H+anti-miR-141-3p group,LPS+Experiment-H+pcDNA3.1 group,LPS+Experiment-H+pcDNA3.1-KLF6 group. Levels of IL-1β and TNF-α were detected by ELISA;Bcl-2 related X protein(Bax)and Kruppel-like factor 6(KLF6)protein expressions were detected by Western blot;cell apoptosis was detected by flow cytometry;miR-141-3p expression was detected by RT-qPCR;targeting relationship betwwen miR-141-3p and KLF6 were detected by luciferase reporter assay.Results:Levels of IL-1β and TNF-α,Bax expression,apoptosis rate in LPS-induced cardiomyocytes were significantly increased,expression of miR-141-3p was significantly decreased,and expression of KLF6 was significantly increased(P<0.05). After treatment with gardenia polysaccharide,levels of IL-1β and TNF-α,Bax expression,apoptosis rate in LPS-induced cardiomyocytes were signifcantly decreased,expression of miR-141-3p was significantly increased,and expression of KLF6 was significantly decreased(P<0.05). Inhibition of miR-141-3p expression and overexpression of KLF6 reversed inhibitory effect of gardenia polysaccharide on H9C2 cells apoptosis and inflammatory factors induced by LPS. miR-141-3p could targeting regulates KLF6 expression.Conclusion:Gardenia polysaccharide can inhibit apoptosis of cardiomyocytes induced by LPS and release of inflammatory factors,which has protective effects on LPS-induced cardiomyocyte injury,whose mechanism may be related to miR-141-3p and KLF6.
作者
刘大朋
李影
孙作乾
宋涛
陈敏
LIU Dapeng;LI Ying;SUN Zuoqian;SONG Tao;CHEN Min(Clinical Medicine Teaching and Research Office,Department of Medical Technology,Zaozhuang Vocational College of Science and Technology,Tengzhou 277599,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第18期2226-2230,共5页
Chinese Journal of Immunology
